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您好，请教下SBI Exoquick TC 试剂盒提取外泌体的相关问题。
小鼠乳腺癌EMT6 cell line，正常培养48小时后换成Exo-free 的10%FBS培养基，培养48-72小时后收集上清，提取外泌体。
1 Collect cell culture medium
2 300g 5min, 2000g 20min
3 Collect the supernatant and pass through 0.22um filter membrane
4 Ultrafiltration tube 100kDa, supernatant 15ml, 4000g 30min, 4 centigrade.
6 Collect the residual liquid on the upper part, about 200ul.
7 Add the Exoquick reagent with the ratio (sample Vs reagent, 5 Vs 1), then inverting or flicking the tube.
8 Incubate overnight, about 16 hours under 4 centigrade
9 Centrifuge 10000g 30min.
10 Collect the pellet and resuspend with PBS.
通常，按网上资料显示，试剂盒提取的外泌体会产生大量沉淀，按此操作后，最终未见沉淀出现，细胞量大概5x107。想请您帮忙分析下原因，不知您是否提取过EMT6细胞系的外泌体，可能是细胞系的原因么，还是实验方案的问题，小鼠的外泌体会和人类的大小存在差别么。谢谢
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|Archiver|手机版|外泌体之家 | exosomes & microvesicles
楼主
发表于 2019-12-3 22:27:42
